Compositions with a biostimulating activity

ABSTRACT

A description follows of compositions with a biostimulating activity comprising at least two components selected from a thiazolidine-4-carboxylic acid, an N-acyl derivative of thiazolidine-4-carboxylic acid and/or an amino acid component consisting of one or more essential amino acid.

The present invention relates to compositions with a biostimulatingactivity and their agronomic use.

It has been known for some time that R-(−)-thiazolidine-4-carboxylicacid [CAS 34592-47-7] (hereafter also indicated as ATC) and some of itsderivatives, in particular, N-acetylthiazolidine-4-carboxylic acid [CAS5025-82-1] (hereafter also indicated as AATC) are used singly ascomponents of compositions with a biostimulating activity tore-establish the correct physiological equilibrium of plants subject tovarious kinds of abiotic or biotic stress.

Different kinds of stress can be caused, for example, by cold, heat,drought, or by exogenous chemical agents such as herbicidal compounds,fungicidal compounds or insecticides, or other kinds of stress can becaused by specific development phases of the vegetables themselves whichcreate particular energy consumption, such as allegation, blossoming,fruit enlargement, etc.

The Applicant has surprisingly found that, by applying compositions witha biostimulating activity consisting of thiazolidine-4-carboxylic acidmixed with N-acyl derivatives of said thiazolidine-4-carboxylic acid,there is a synergic activity between these compounds.

The Applicant has also surprisingly found that, by applying compositionswith a biostimulating activity consisting of thiazolidine-4-carboxylicacid and/or an N-acyl derivative of thiazolidine-4-carboxylic acidand/or an amino acid component consisting of one or more essential aminoacids, there is an amplification of the biostimulating effect of theabove composition, or a synergic activity when said amino acid componentproves to have a biostimulating activity.

An object of the present invention therefore relates to compositionscomprising at least two components selected fromthiazolidine-4-carboxylic acid, an N-acyl derivative ofthiazolidine-4-carboxylic acid and an amino acid component consisting ofone or more essential amino acids.

A further object of the present invention relates to the use ofcompositions comprising at least two components selected fromthiazolidine-4-carboxylic acid, an N-acyl derivative ofthiazolidine-4-carboxylic acid and an amino acid component consisting ofone or more essential amino acids, as biostimulants in the agronomicfield.

In particular, the N-acyl derivative of thiazolidine-4-carboxylic acidis preferably selected from N-acetyl-thiazolidine-4-carboxylic acid,N-propanoyl-thiazolidine-4-carboxylic acid orN-butanoyl-thiazolidine-4-carboxylic acid.

Preferred compositions according to the present invention comprisethiazolidine-4-carboxylic acid and N-acetyl-thiazolidine-4-carboxylicacid or thiazolidine-4-carboxylic acid and an amino acid componentconsisting of one or more essential amino acids orN-acetyl-thiazolidine-4-carboxylic acid and an amino acid componentconsisting of one or more essential amino acids.

A further preferred composition according to the present inventioncomprises thiazolidine-4-carboxylic acid andN-acetyl-thiazolidine-4-carboxylic acid and an amino acid componentconsisting of one or more essential amino acids.

The N-acyl derivatives of thiazolidine-4-carboxylic acid can be presentin acid form or in salified form such as salts of sodium, potassium,magnesium, copper, iron, manganese, calcium, or of another inorganiccation.

Furthermore, the N-acyl derivatives of thiazolidine-4-carboxylic acidcan be present in salified form such as ammonium salts, or salified withan organic amine, such as triethylamine, cyclohexylamine,trimethylamine, triethanolamine, or with an amine group of a basic aminoacid such as Glutamic acid, or ornithine.

The amino acid component consisting of one or more essential amino acidsis preferably selected from proline and/or glutamic acid and/orglutamine. In particular the proline and/or glutamic acid and/orglutamine can be in purified form or as complex mixtures.

The amino acid components can consist of amino acids in purified form orconsist of complex mixtures deriving from partial or total hydrolysis ofbiological matrixes containing protein fractions.

These amino acids or mixtures of essential amino acids can therefore befree amino acids or proteinic amino acids, which, within the context ofthe present invention, are considered as being precursors of free aminoacids.

Said proteinic amino acids can consequently be incorporated indipeptides, tripeptides or oligopeptides or peptides with differentmolecular weights.

The amino acid component according to the present invention cantherefore consist of hydrolyzed proteinic products obtained from naturalsubstrates.

These hydrolyzed products can have varying degrees of hydrolysis andwill therefore have free amino acids and/or dipeptides, tripeptides oroligopeptides with varying molecular weights. Said hydrolyzed proteinicproducts preferably contain at least 5% of free amino acids with respectto the total amino acid content, and will preferably have an overallminimum content of total amino acids (free or proteinic) of 25% w/w.

The hydrolyzed proteinic products are preferably hydrolyzed proteinicproducts of bovine epithelium or vegetable matrixes.

Said hydrolyzed product will preferably contain essential amino acids,such as proline, glutamic acid or glutamate, or glutamine and can haveits own biostimulating activity on numerous crops of economic interest.

The use of the composition according to the present invention isparticularly advantageous specifically as a result of the synergiceffect observed in the application of the compositions with abiostimulating activity comprising at least two components selected fromthiazolidine-4-carboxylic acid, an N-acyl derivative ofthiazolidine-4-carboxylic acid and an amino acid component consisting ofone or more essential amino acids, when said amino acid component provesto have its own biostimulating activity.

A further advantage consists in the amplification of the biostimulatingeffect observed in the application of the compositions with abiostimulating activity comprising at least two components selected fromthiazolidine-4-carboxylic acid, an N-acyl derivative ofthiazolidine-4-carboxylic acid and an amino acid component consisting ofone or more essential amino acids, when said amino acid component doesnot have its own biostimulating activity.

In agronomic practice, the compositions object of the present inventioncan conveniently contain additives, dispersants, wetting agents and anyother product known in the art for favouring the application of activesubstances to vegetables.

An object of the present invention therefore also relates to a methodfor the biostimulation of agricultural crops by the application of thecompositions according to the present invention, alone or with othercompounds having the same or a different activity.

The application of the compositions object of the present invention iseffected on the whole plant or a part of it, by application on theaerial part and/or on the hypogeum, for example by dispersion orspraying, on the leaves or via the roots.

Said thiazolidine-4-carboxylic and N-acyl-thiazolidine-4-carboxylicacids and the amino acid components are generally commercial productseasily available on the market. The hydrolyzed proteinic product frombovine epithelium, sold in Italy and in many other countries in theworld under the trade mark of Siapton® and the hydrolyzed product basedon vegetables (soybean) called Fertimin® are examples of a complex aminoacid mixture.

The following examples are provided for merely illustrative purpose andin no way intend to limit the scope of the present invention. In thefollowing examples the synergic effect of the components of the mixtureis revealed by comparing the experimental data with the theoreticalefficacy of the mixture, object of the invention, calculated accordingto the Limpel formula (“Pesticide Science” (1987), vol. 19, pages309-315):E=x+y−(xy/100)wherein:

-   -   E is the biological activity expected, in the absence of        synergic effects, from a mixture obtained by mixing x g of        compound X with y g of compound Y;    -   x is the activity of the compound X when used alone at a dose of        x g;    -   y is the activity of the compound Y when used alone at a dose of        y g.

When the biological activity experimentally found is higher than thevalue of E, said activity should be considered as a synergic effect.

EXAMPLE 1

Preparation of compound with a biostimulating activity for thesubsequent experiments.

An amount of 50 g of each compound under examination(thiazolidine-4-carboxylic acid (compound I) andN-acyl-thiazolidine-4-carboxylic acid (compound II)) were dispersed inwater (800 cc) and vigorously stirred. An excess of 5%, with respect tothe stoichiometric quantity, of sodium bicarbonate, was then added inportions. 150 g of a non-ionic surface-active agent were added to favourapplication to the leaves. The whole mixture was brought to a volume of1 liter by the addition of water, in order to obtain solutions of eachcompound with a concentration of 50 g/liter.

EXAMPLE 2

Assessment of the biostimulating activity of compound (I), compound (II)and their synergic mixture.

Seedlings of courgettes cv. Diamone, sown and cultivated in a hotgreenhouse, were transplanted in a field (May 11, Leggiuno,(Varese)—Italy).

The theses were randomly arranged in lots with four repetitions for eachthesis. Each lot consisted of six plants in rows situated at a distanceof 1 meter from each other, and at a distance of 1 meter from each otherin the same row.

Weeding was performed before the transplant, using 1 kg/ha oftrifluralin (volume 550 l/ha).

At the moment transplanting, the seedlings have a common degree ofvegetative development.

In order to minimize transplant stress, the seedlings planted weretreated respectively with:

-   -   Thesis A: with compound (I) (ATC) at a dose of 5 g/hl;    -   Thesis B: with compound (I) (ATC) at a dose of 10 g/hl;    -   Thesis C: with compound (II) (AATC) at a dose of 5 g/hl;    -   Thesis D: with compound (II) (AATC) at a dose of 10 g/hl;    -   Thesis E: with compound (I) (ATC) at a dose of 5 g/hl in a        mixture with compound (II) (AATC) at a dose of 5 g/hl;    -   Thesis F: no biostimulating treatment.

Said treatment was subsequently repeated every 10 days, in a preventivemanner, in order to oppose any possible thermal or water stress orstress due to vegetative development. Harvesting was effected from June11 to August 27, and the yield increases indicated in Table 1, wereregistered in the treated theses (A, B, C, D, E), with respect to thenon-treated Thesis F. The yields are obtained from the average amount offruit obtained in each lot during the above-mentioned harvesting period(June 11-August 27). TABLE 1 THESIS C D E A B (5 g (10 g (5 g AATC/ F (5g (10 g AATC/ AATC/ hl + (not ATC/h) ATC/hl) hl) hl) 5 g ATC/hl)treated) Fruit per 100 105 102 109 115 97 lot Increase +3.1% +8.2% +5.2%+12.4% +18.6% — (%)

From the experimental results obtained, the expected activity (E) forthe mixture of 5 g of compound I plus 5 g of compound II (Thesis E),calculated by applying to said experimental results the above-mentionedLimpel formula, is:E=(3.1+8.2)−(3.1·8.2)/100=11%

The ratio between the experimental data and the expected activity provesto be higher than 1, indicating a synergy of effects:18.6/11=1.69

EXAMPLE 3

Determination of the amplification of the biostimulating effect obtainedby applying compound (I) in the presence of a solution of proline.

Seedlings of peppers cv. Pathos were transplanted in a tunnel (April 16,AT).

The lots were distributed according to the random block scheme with 5repetitions per thesis. Each repetition consisted of 15 plants.

At the moment of transplanting, the seedlings have a common degree ofvegetative development.

In order to minimize transplant stress, the seedlings planted weretreated respectively with:

-   -   Thesis A: with compound (I) (ATC) at a dose of 10 g/hl;    -   Thesis B: with compound (I) (ATC) at a dose of 10 g/hl, an        aqueous solution of proline at a dose of 100 g/hl;    -   Thesis C: with compound (II) (AATC) at a dose of 10 g/hl;    -   Thesis D: with compound (II) (AATC) at a dose of 10 g/hl, an        aqueous solution of proline at a dose of 100 g/hl;    -   Thesis E: no biostimulating treatment.

Said treatment was subsequently repeated every 10 days, in a preventivemanner, in order to oppose any possible thermal or water stress orstress due to vegetative development.

Harvesting was effected from July 29 for the following 30 days, and aproduction increase was registered in theses B and D, with respect totheses A and C, as indicated in Table 2. The yields are obtained fromthe average production obtained during the above harvesting period (July29-August 29). TABLE 2 THESIS B D (10 g (10 g A ATC/hl + C AATC/hl + E(10 g 100 g (10 g 100 g (not ATC/hl) Proline/hl) AATC/hl) Proline/hl)treated) Berries kg 25.80 27.80 26.40 28.20 24.70 per lot Increase +4.5%+12.6% +6.9% +17.0% — (%)

As it is known that the proline solutions do not have their ownbiostimulating activity, this amplification is attributed to a synergicinteraction of the proline itself on the biostimulating activity ofcompounds (I) and (II).

EXAMPLE 4

Determination of the synergic effect obtained in the application of amixture based on compound (I) and compound (II) to which a mixture isadded, containing essential amino acids, consisting of a hydrolyzedproteinic product obtained from bovine epithelium, with a knownbiostimulating activity.

Tomato seedlings cv. Arletta, sown and cultivated in a greenhouse, weretransplanted in a field (May 11, Leggiuno (VA)—Italy), after weedingwith trifluralin (1 kg/ha).

The theses were arranged in random blocks. Each lot consisted of 10plants at a distance of 50 cm from each other in rows 80 cm apart.

At the moment of transplanting, the seedlings have a common degree ofvegetative development.

In order to minimize transplant stress, the seedlings planted weretreated respectively with:

-   -   Thesis A: with compound (I) (ATC) at a dose of 5 g/hl mixed with        compound (II) (AATC) at a dose of 5 g/hl;    -   Thesis B: with a hydrolyzed proteinic product of an animal        origin (compound III), at a dose of 500 g/hl;    -   Thesis C: with compound (I) (ATC) at a dose of 5 g/hl mixed with        compound (II) (AATC) at a dose of 5 g/hl mixed with a hydrolysed        proteinic product (compound III) at a dose of 500 g/hl;    -   Thesis D: no biostimulating treatment.

Said treatment was subsequently repeated every 15 days, in a preventivemanner, in order to oppose any possible thermal or water stress orstress due to vegetative development.

In particular, the compound (III) used in this test, for the sole andnon-limiting purpose of illustrating the behaviour of a hydrolyzedproteinic product, is a sample of Siapton®, a hydrolyzed product ofbovine epithelium, characterized by an overall content of about 700 g ofamino acids (free and proteinic) per liter of solution, of which about13% w/w consists of proline, about 12% of glutamic acid, about 9.5% of4-hydroxyproline.

Harvesting was effected from July 16 to September 9, and the yieldincreases indicated in Table 3, were registered in the theses treated(A, B, C) with respect to the non-treated Theses D. The yields areobtained from the average amount and average weight of the fruitobtained per lot, during the above harvesting period (July 16-September9). TABLE 3 THESIS C B (5 g C A Com- AATC/hl + 5 g Expected (5 g poundATC/hl) + Activity D AATC/hl + III Compound III according (not 5 gATC/hl) (XX g/hl) (500 g/hl) to Limpel treated) Berries 178 173 193 —161 per lot Increase +10.6%  +7.5% +19.9% +18.0% — (%) Berries 22.1021.40 25.10 18.60 kg/lot Increase +18.8% +15.1% +34.9% +33.8% — (%)

The expected values applying the Limpel formula are lower than thoseobtained experimentally showing a synergy of effects.

EXAMPLE 5

Determination of the synergic effect obtained in the application of amixture based on compound (I) and compound (II) to which a mixture isadded, containing essential amino acids, consisting of a hydrolyzedproteinic product of a vegetable origin (compound IV).

Aubergine seedlings cv. Violetta Lunga, sown and cultivated in a hotgreenhouse, were transplanted in the middle of a field on May 14(Leggiuno (VA)—Italy), after weeding with trifluralin (1 kg/ha).

The theses were distributed according to 5 random blocks. Each lotconsisted of 8 plants at a distance of 40 cm from each other on eachrow. The distance between the rows was 1 m.

At the moment of transplanting, the seedlings have a common degree ofvegetative development.

In order to minimize transplant stress, the seedlings planted weretreated respectively with:

-   -   Thesis A: with compound (I) (ATC) at a dose of 5 g/hl mixed with        compound (II) (AATC) at a dose of 5 g/hl;    -   Thesis B: with a hydrolyzed proteinic product (compound IV) with        a minimum content of proline of 1.5% (meaning overall free or        proteinic proline) at a dose of 900 g/hl;    -   Thesis C: with compound (I) (ATC) at a dose of 5 g/hl mixed with        compound (II) (AATC) at a dose of 5 g/hl mixed with a hydrolysed        proteinic product (compound IV) at a dose of 600 g/hl;    -   Thesis D: no biostimulating treatment.

Said treatment was subsequently repeated every 10 days, in a preventivemanner, in order to oppose any possible thermal or water stress orstress due to vegetative development.

In particular, the compound (IV) used in this test, for the sole andnon-limiting purpose of illustrating the behaviour of a hydrolyzedproteinic product, is a hydrolyzed proteinic product obtained fromsoybean and characterized by a minimum content of proline of about 1.5%,a minimum content of glutamic acid of about 5% and the total absence of4-hydroxyproline.

Harvesting was effected from August 1 to September 12, and the yieldincreases indicated in Table 4, were registered in the theses treated(A, B, C) with respect to the non-treated Theses D. The yields areobtained from the average amount of fruit obtained for each lot, duringthe above harvesting period (August 1-September 12). TABLE 4 THESIS C (5g B AATC/hl) + C A Com- 5 g ATC/hl) + Expected (5 g pound CompoundActivity D AATC/hl + IV IV according to (not 5 g ATC/hl) (600 g/hl) (600g/ha) Limpel treated) Berries 28.1 26.7 30.4 — 25.8 per lot Increase+8.9% +3.5% +17.8% +12.4% — (%)

The expected values applying the Limpel formula are lower than thoseobtained experimentally showing a synergy of effects.

1. Compositions comprising at least two components selected from thiazolidine-4-carboxylic acid, an N-acyl derivative of thiazolidine-4-carboxylic acid and an amino acid component consisting of one or more essential amino acids.
 2. The compositions according to claim 1, characterized in that the N-acyl derivative of thiazolidine-4-carboxylic acid is selected from N-acetyl-thiazolidine-4-carboxylic acid, N-propanoyl-thiazolidine-4-carboxylic acid or N-butanoyl-thiazolidine-4-carboxylic acid.
 3. The compositions according to claim 1, characterized in that they comprise thiazolidine-4-carboxylic acid and N-acetyl-thiazolidine-4-carboxylic acid.
 4. The compositions according to claim 1, characterized in that they comprise thiazolidine-4-carboxylic acid and an amino acid component consisting of one or more essential amino acids.
 5. The compositions according to claim 1, characterized in that they comprise N-acetyl-thiazolidine-4-carboxylic acid and an amino acid component consisting of one or more essential amino acids.
 6. The compositions according to claim 1, characterized in that they comprise thiazolidine-4-carboxylic acid, N-acetyl-thiazolidine-4-carboxylic acid and an amino acid component consisting of one or more essential amino acids.
 7. The compositions according to claim 1, characterized in that the N-acyl derivatives of thiazolidine-4-carboxylic acid are present in acid form or in salified form.
 8. The compositions according to claim 7, characterized in that the N-acyl derivatives of thiazolidine-4-carboxylic acid are present in salified form as salts of sodium, potassium, magnesium, copper, iron, manganese, calcium, or another inorganic cation.
 9. The compositions according to claim 7, characterized in that the N-acyl derivatives of thiazolidine-4-carboxylic acid are present in salified form as ammonium salts, or salified with an organic amine such as triethylamine, cyclohexylamine, trimethylamine, triethanolamine, or with the amine group of a basic amino acid such as glutamic acid or ornithine.
 10. The compositions according to claim 1, characterized in that the amino acid component consisting of one or more essential amino acids is selected from proline and/or glutamic acid and/or glutamine.
 11. The compositions according to claim 10, characterized in that proline and/or glutamic acid and/or glutamine are in purified form or are complex mixtures.
 12. The compositions according to claim 1, characterized in that the essential amino acids are in purified form or consist of complex mixtures deriving from partial or total hydrolyses of biological matrixes containing proteinic fractions.
 13. The compositions according to claim 1, characterized in that the amino acids or mixtures of essential amino acids consist of free amino acids or proteinic amino acids, precursors of free amino acids.
 14. The compositions according to claim 1, characterized in that the amino acid component consists of hydrolyzed proteinic products obtained from natural substrates.
 15. The compositions according to claim 14, characterized in that the hydrolyzed proteinic products contain at least 5% of free amino acids with respect to the total content of amino acids.
 16. The compositions according to claim 14, characterized in that the hydrolyzed proteinic products have an overall minimum content of total amino acids (free or proteinic) of 25% w/w.
 17. The compositions according to claim 14, characterized in that the hydrolyzed proteinic products are hydrolyzed proteinic products of bovine epithelium.
 18. The compositions according to claim 14, characterized in that the hydrolyzed proteinic products are hydrolyzed proteinic products of vegetable matrixes.
 19. The compositions according to any of the previous claims, characterized in that they comprise further additives, dispersants and/or wetting agents.
 20. A method for biostimulating agricultural crops by the application of the compositions according to claim 1, alone or with other compounds having an analogous or different activity.
 21. The method according to claim 20, characterized in that the application takes place on the whole plant or on a part of the plant, by means of applications on the aerial part and/or on the hypogeum, for example, by dispersion or spraying, either on the leaves or via the roots.
 22. (canceled) 